中国中药杂志

2021, v.46(24) 6520-6529

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粉防己碱对胶质母细胞瘤细胞增殖、迁移和侵袭的影响及其机制研究
Effects of tetrandrine on proliferation, migration, and invasion of glioblastoma cells

鲁鑫宇;王忠泽;万思成;赵二虎;崔红娟;
LU Xin-yu;WANG Zhong-ze;WAN Si-cheng;ZHAO Er-hu;CUI Hong-juan;College of Sericulture, Textile and Biomass Sciences, State Key Laboratory of Silkworm Genome Biology, Southwest University;Medical Research Institute, Southwest University;Westa College, Southwest University;

摘要(Abstract):

胶质母细胞瘤是常见的颅内原发恶性肿瘤,患者生存质量不佳且复发率高,化学药物治疗是其治疗至关重要的一部分。粉防己碱(tetrandrine, Tet)是一种从中药粉防己根部提取出来的活性成分,多项研究表明其具有包括抗肿瘤在内的广泛药理作用,但鲜有其在胶质瘤相关研究的报道。该研究以胶质母细胞瘤细胞LN229为实验对象,采用MTT法和BrdU实验检测Tet对胶质母细胞瘤细胞增殖能力的影响,流式细胞仪分析胶质母细胞瘤细胞在Tet作用后细胞周期和凋亡的变化,平板克隆实验和软琼脂克隆形成实验研究Tet对肿瘤细胞克隆形成能力的影响,划痕实验和Transwell实验研究Tet对肿瘤细胞迁移和侵袭能力的调节,Western blot法分析Tet影响细胞增殖、迁移和侵袭的分子机制。在Tet的作用下,MTT实验、BrdU实验结果表明胶质瘤细胞系LN229的增殖活力受到抑制并且具有时间和剂量效应;平板克隆实验和Soft Agar实验显示Tet降低了LN229细胞体外克隆形成能力;流式细胞术实验说明Tet将细胞阻滞在G_1期并促进了细胞凋亡;划痕实验和Transwell实验证明Tet可以抑制LN229细胞的迁移和侵袭;Western blot结果显示Tet可以下调CDK2、CDK6、cyclin D1、cyclin E1、snail、slug、vimentin和N-cadherin的水平,并且上调E-cadherin的水平。由此推测Tet对胶质母细胞瘤细胞LN229的增殖、迁移以及侵袭能力具有一定的抑制作用,这种抑制作用可能与其参与调节c-Myc/p27轴、snail信号通路有关。
Glioblastoma is the most common intracranial primary malignant tumor, which leads to the poor quality of life of patients and has a high recurrence rate. Chemotherapy is a vital part in the treatment of this disease. Tetrandrine(Tet) is an active ingredient extracted from the root of the Chinese medicinal plant Stephania tetrandra, which has been proved with a wide range of pharmacological effects including anti-tumor. However, there are few studies regarding the effect of Tet on glioma. In this study, MTT and BrdU assays were employed to detect the effect of Tet on the proliferation of LN229 glioblastoma cells; flow cytometry was used to analyze the cycle distribution and apoptosis; plate cloning assay and soft agar colony formation assay were performed to study the colony formation ability of LN229 cells exposed to Tet; scratch assay and Transwell assay were conducted to detect the ability of migration and invasion; Western blot was adopted to the exploration of the molecular mechanism. The MTT and BrdU assays showed that Tet inhibited the proliferation of LN229 cells in a time-and dose-dependent manner. The plate cloning assay and soft agar colony formation assay showed that Tet weakened the colony formation of LN229 cells in vitro; cytometry assay showed that Tet blocked cells in the G_1 phase and promoted cell apoptosis; scratch and Transwell assays proved that Tet inhibited the migration and invasion of LN229 cells; Western blot results showed that Tet down-regulated the expression levels of CDK2, CDK6, cyclin D1, cyclin E1, snail, slug, vimentin, and N-cadherin, while up-regulated the level of E-cadherin. The results indicate that Tet has a certain inhibitory effect on the proliferation, migration, and invasion of LN229 glioblastoma cells, and such effect may be related to the participation of Tet in the regulation of c-Myc/p27 axis and snail signaling pathway.

关键词(KeyWords): 粉防己碱;胶质母细胞瘤;增殖;迁移;侵袭
tetrandrine;glioblastoma;proliferation;migration;invasion

Abstract:

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基金项目(Foundation): 国家自然科学基金项目(81972357);; 重庆市大学生创新创业训练计划项目(S202010635071)

作者(Author): 鲁鑫宇;王忠泽;万思成;赵二虎;崔红娟;
LU Xin-yu;WANG Zhong-ze;WAN Si-cheng;ZHAO Er-hu;CUI Hong-juan;College of Sericulture, Textile and Biomass Sciences, State Key Laboratory of Silkworm Genome Biology, Southwest University;Medical Research Institute, Southwest University;Westa College, Southwest University;

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